Fig. 4

Sox9 overexpression promoted hASC chondrogenic differentiation by blocking the effect of miR-539-3p mimic. hASCs were transfected with miR-539-3p mimic or/and pcDNA-Sox9, which were cultured in chondrogenic differentiation medium. hASCs cultured in high-glucose DMEM with 1% fetal bovine serum (FBS) were used as control. Gene expression levels of COL2A1 and ACAN in hASCs or CD-ASCs were determined by RT-qPCR (A and B) and western blot (C–E). β-actin expression levels were detected as an endogenous control. ^**P < 0.01 (VS. ASCs), ^##P < 0.01 (VS. ASCs), ^&P < 0.05 (VS. miR-539-3p mimic), ^&&P < 0.05 (VS. miR-539-3p mimic), ^$$P < 0.01 (VS.pcDNA-Sox9). (I) Immunofluorescence assay of Type II collagen. Type II collagen fluorescence (green) and DAPI (blue). Scale bar: 100 μm and 50 μm. The experiment was repeated at least 3 times independently. The data are presented as means ± standard deviation (SD)